ATP-dependent force generation and membrane scission by ESCRT-III and Vps4

Johannes Schöneberg; Mark Remec Pavlin; Shannon Yan; Maurizio Righini; Il-Hyung Lee; Lars-Anders Carlson; Amir Houshang Bahrami; Daniel H Goldman; Xuefeng Ren; Gerhard Hummer; Carlos Bustamante; James H Hurley

doi:10.1126/science.aat1839

ATP-dependent force generation and membrane scission by ESCRT-III and Vps4

Science. 2018 Dec 21;362(6421):1423-1428. doi: 10.1126/science.aat1839.

Authors

Johannes Schöneberg^{1

2

3}, Mark Remec Pavlin^#^{2

4}, Shannon Yan^#^{1

2}, Maurizio Righini⁵, Il-Hyung Lee^{1

2}, Lars-Anders Carlson^{1

2}, Amir Houshang Bahrami³, Daniel H Goldman^{2

6

5}, Xuefeng Ren^{1

2}, Gerhard Hummer^{3

7}, Carlos Bustamante^{8

2

4

6

5

9

10}, James H Hurley^{8

2

4

10}

Affiliations

¹ Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
² California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA 94720, USA.
³ Department of Theoretical Biophysics, Max Planck Institute of Biophysics, 60438 Frankfurt am Main, Germany.
⁴ Graduate Group in Biophysics, University of California, Berkeley, Berkeley, CA 94720, USA.
⁵ Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720, USA.
⁶ Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA 94720, USA.
⁷ Institute of Biophysics, Goethe University, Frankfurt/M, Germany.
⁸ Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA. jimhurley@berkeley.edu carlosb@berkeley.edu.
⁹ Department of Physics, University of California, Berkeley, Berkeley, CA 94720, USA.
¹⁰ Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.

^# Contributed equally.

Abstract

The endosomal sorting complexes required for transport (ESCRTs) catalyze reverse-topology scission from the inner face of membrane necks in HIV budding, multivesicular endosome biogenesis, cytokinesis, and other pathways. We encapsulated ESCRT-III subunits Snf7, Vps24, and Vps2 and the AAA+ ATPase (adenosine triphosphatase) Vps4 in giant vesicles from which membrane nanotubes reflecting the correct topology of scission could be pulled. Upon ATP release by photo-uncaging, this system generated forces within the nanotubes that led to membrane scission in a manner dependent upon Vps4 catalytic activity and Vps4 coupling to the ESCRT-III proteins. Imaging of scission revealed Snf7 and Vps4 puncta within nanotubes whose presence followed ATP release, correlated with force generation and nanotube constriction, and preceded scission. These observations directly verify long-standing predictions that ATP-hydrolyzing assemblies of ESCRT-III and Vps4 sever membranes.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Adenosine Triphosphatases / chemistry
Adenosine Triphosphatases / metabolism*
Adenosine Triphosphate / metabolism*
Biocatalysis
Cell Membrane / metabolism*
Cell Membrane / ultrastructure
Endosomal Sorting Complexes Required for Transport / chemistry
Endosomal Sorting Complexes Required for Transport / metabolism*
Hydrolysis
Nanotubes
Saccharomyces cerevisiae Proteins / chemistry
Saccharomyces cerevisiae Proteins / metabolism*
Unilamellar Liposomes

Substances

DID4 protein, S cerevisiae
Endosomal Sorting Complexes Required for Transport
SNF7 protein, S cerevisiae
Saccharomyces cerevisiae Proteins
Unilamellar Liposomes
VPS4 protein, S cerevisiae
Vps24 protein, S cerevisiae
Adenosine Triphosphate
Adenosine Triphosphatases

Abstract

Publication types

MeSH terms

Substances

Grants and funding